Cloning, expression and characterization of recombinant CagA protein of Helicobacter pylori using monoclonal antibodies: Its potential in diagnostics

Barik A. Salih*, Cebrail Karakus, Zeynep Ulupinar, Fahri Akbas, Merve Uslu, Duygu Yazici, Bora Kazim Bolek, Nizamettin Bayyurt, Cansel Turkay

*Bu çalışma için yazışmadan sorumlu yazar

Araştırma sonucu: ???type-name???Makalebilirkişi

2 Atıf (Scopus)

Özet

Helicobacter pylori CagA protein plays an important role in the severity of the gastric diseases. Our aims were to clone the cagA 5′- conserved region of the gene, characterize the recombinant CagA (rCagA) protein by monoclonal antibodies (mAbs) and to use this protein for the detection of anti-CagA antibodies by an ELISA test. Our developed rCagA protein (67 kDa) showed an amino acid sequence homology of 83% and 80% with Western and East Asian type strains respectively. Two anti-rCagA (BS-53, CK-02) mAbs and 2 additional rCagA proteins of smaller sizes (60 kDa, 28 kDa) were developed for epitope determination. The BS-53 mAb recognized all 3 rCagA proteins while CK-02 mAb recognized only 2 of them indicating recognition of different epitopes. An in-house indirect ELISA using rCagA was developed to detect anti-CagA antibodies in sera of 59 patients. The ELISA results obtained when compared to those of the PCR gave a sensitivity, specificity and accuracy of 81%, 100% and 88% respectively. We have developed for the first time: a rCagA protein that showed high sequence homology with both Western and East Asian type strains and an indirect ELISA of high performance which can be used to detect anti-CagA antibodies in sera of infected patients worldwide.

Orijinal dilİngilizce
Sayfa (başlangıç-bitiş)26-31
Sayfa sayısı6
DergiBiologicals
Hacim68
DOI'lar
Yayın durumuYayınlandı - Kas 2020

Bibliyografik not

Publisher Copyright:
© 2020 International Alliance for Biological Standardization

Finansman

This study was supported by the grant of the Scientific and Technological Research Council of Turkey (TÜBİTAK) (No: 111T370 ). The authors would like to thank Dr. Fatima Yucel, Dr. Esin Akcael, Dr. Hivda Polat and Harun Kocaaga for their technical assistance.

FinansörlerFinansör numarası
TÜBİTAK
Türkiye Bilimsel ve Teknolojik Araştirma Kurumu111T370

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